DATA SHEET

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PRODUCT NAME: Monoclonal mouse anti-human MUC1 glycoprotein
CATALOGUE #: 4M40

MAbs:
CM1, X19, M9E7, M4H2, M8C9, M10G4, M10H6, M2F1, M2C5, M2E104, M3A106
Hybridoma clones have been derived from hybridization of X63-Ag8-653 myeloma cells with spleen cells of Balb/c mice immunized with high molecular weight (>300 kDa) glycoprotein from human milk-fat globule molecule.

Specificity:
MAb CM1 - peptide tandem repeat in MUC1 core. Similar to DF3 MAb (Centocor). Affinity more than 2 x  10⁹ M^-1 . MAb X19 - peptide repeat RPAP region
MAbs M9E7, M8C9, M10G4, M10H6: Affinity > 1 x 10⁸ for all MAbs. MAbs bind with high efficiency with different regions of GVTSAPDTRPAPGSTAPPAHGVTSA synthetic peptide spanning the one repeat of VNTR extracellular portion of MUC1 molecule. They react with VNTR5 and VNTR20 recombinant unglycosylated fragments of MUC1 protein and underglycosylated MUC1 prepared from tumor fluids or as a result of chemical treatment of human milk MUC1. These MAbs do not recognize natural MUC1 protein isolated from human milk by affinity purification on carbohydrate epitope specific MAbs and belong to MAb cluster 1-4 according to ISOMB TD-4 classification. No cross-reactivity with egg white avidin.
MAbs M4H2, M2F1, M2C5, M2E104 and M3A106: Affinity >1 X 10⁸ for all MAbs. MAbs bind with high efficiency with different epitopes within the VNTR tandem repeat peptide region of MUC1 molecule, which have different conformations and peptide- carbohydrate compositions. MAb M4H2 reacts with VNTR20 recombinant unglycosylated fragment of MUC1 protein (and very weakly with monomeric MUC1 peptide), with deglycosylated and underglycosylated natural MUC1 (but not with completely glycosylated natural milk MUC1). MAbs M2F1, M2C5, M2E104, M3A106 efficiently recognize underglycosylated and natural MUC1 protein isolated from human milk by affinity chromatography. All these MAbs belong to MAb clusters 6 and 7 according to ISOMB TD-4 classification. Non crossreactive with normal serum proteins and human tissues of non-epithelial
origin.

MAb isotypes:
IgG for MAbs M9E7, M4H2, M8C9, M10G4, M10H6, M4H2, M2F1, M2C5, M2E104 and M3A106
IgG1 for MAbs CM1 and X19

Applications:
Immunohistochemical and immunofluorescent detection of neoplastic breast epithelium. Antibodies are working on frozen tissue sections. Construction of immunometric assay for MUC determination.
MAbs M9E7, M4H2, M8C9, M10G4, M10H6 can be used for construction of highly effective ELISA tests for detection and measurements of unglycosylated and underglycosylated tumor-associated MUC1 mucins in human serum. They may be used as solid-phase reagents for MUC1 isoforms capture
(a) in conjunction with the same MAbs enzyme-conjugated vertions (detection of unglycosylated
MUC1), or
(b) with enzyme-conjugated MAbs M2C5, M2E10 or M3A10 specific to carbohydrate portions of MUC1 (detection of underglycosylated MUC1).
MAbs M4H2, M2F1, M2C5, M2E104, M3A106 can be used for immunohistochemical staining of paraffin and frozen sections of breast, lung and ovarian tumor tissues. They may be used for staining of paraffin-embedded tissue sections fixed in neutral-buffered formalin and is compatible with commonly used histological fixatives. Revealed MUC1 epitopes are abundantly present as on the surface of growing tumor cell lines, so as on the surface and into the cytoplasm of malignant cells of epithelial origin. These MAbs can be used in the form of enzyme conjugates for development in ELISA sandwich assays of under-glycosylated tumor-associated MUC1 mucins in human serum. They may be used in conjunction with MAbs M9E7, M4H2, M8C9, M10G4 and M10H6 specific to unglycosylated VNTR region of MUC1 as capture reagents.

Purification:
Chromatography on protein A Sepharose

Presentation:
PBS, pH 7.4, 0.1 % sodium azide (NaN3)

Storage:
+ 4 °C

Material safety note:
This product is sold as an antibody preparation for research use only. Standard Laboratory Practices should be followed when handling this material.
Contains sodium azide (0.1 %) as preservative. Although the amount of sodium azide is very small appropriate care must be taken when handling this product.